Assessing memory representations in freely-behaving animals with calcium imaging via miniscopes

对于这项RTI拨款，我们请求获得“最先进的”加州大学洛杉矶分校微型显微镜(版本4)的支持，以便在自由行为的啮齿动物中进行体内钙成像。这些微型望远镜将是莱斯布里奇大学(UofL)加拿大行为神经科学中心(CCBN)新出现的脑成像核心的重要补充。建议的成像设备将由至少9名来自CCBN的NSERC资助的系统神经科学家和一名来自加州大学洛杉矶分校生物科学系的用户使用。所有用户都将获得使用该设备的权限和技术支持。需要加州大学洛杉矶分校的系统有几个原因：1)在动物自由行为的同时，能够在大脑的某个区域进行钙成像，如海马体(HPC)，这是一项重大进步，可以增加我们可以评估的功能范围；2)它具有任务灵活性；3)可以在不同的大脑区域使用；4)它对用户友好；5)它是这项技术最负盛名的系统。 加州大学洛杉矶分校的成像系统在我们的几个研究项目中将是必不可少的，包括：1)使用相同或不同的空间信息评估快速新学习过程中形成的HPC表征，我们假设这些空间信息的内容和潜在的神经生物学机制将不同。微型显微镜将被用来确定在变化的条件下表示如何不同；2)调查昼夜节律扰动对HPC功能的影响。迷你望远镜将被用来评估各种形式的昼夜节律扰动(即轮班工作)对HPC表征的影响；3)哺乳动物大脑中的记忆功能似乎是由调节不同记忆功能的神经网络组织的，在这个组织中，正常的思维和行为来自这些系统之间的合作和竞争相互作用。我们将使用微型望远镜在不同的实验条件下对皮层下和皮质区域的活动进行成像，以探索多个记忆网络之间相互作用的机制。 使用UCLA-V4添加体内钙成像将对我们的受训人员非常有益，因为：1)由于各种原因，光学成像方法比传统的电生理方法越来越受欢迎，包括可以收集大量数据，但操作和维护设备所需的技术专业知识较少；2)理解从微型显微镜收集的大量数据需要计算技能，我们的受训人员将获得这些技能；以及3)提高生成出版物的质量。所有这些方面都将对我们的学员有利，因为他们在神经科学领域寻求竞争空间。 我们将使用这项技术来推进我们的知识，即行为是如何通过接触特定事件来塑造的，负责的神经网络，以及支持这些功能的生化机制。我们的研究将使所有加拿大人受益，因为我们扩大了对正常和不正常行为的理解。

For this RTI grant, we request support to obtain “state of the art” UCLA miniscopes (version 4) to conduct in-vivo calcium imaging in freely behaving rodents. These miniscopes will be a crucial addition to our emerging Brain Imaging Core at the Canadian Centre for Behavioural Neuroscience (CCBN) at the University of Lethbridge (UofL). The proposed imaging equipment will be used by at least 9 NSERC-funded system neuroscientists from the CCBN, and a user from the Department of Biological Sciences at the UofL. All users will have access and technical support to use this equipment. The UCLA system is requested for several reasons: 1) ability to do calcium imaging in a brain area, like hippocampus (HPC), while the animal is freely-behaving is a major advancement that increases the range of functions we can assess; 2) it is task-flexible; 3) can be used in different brain regions; 4) it is user friendly; and 5) it is the most reputable system for this technique. The UCLA imaging system will be essential in several of our research programs including: 1) assess HPC representations formed during rapid new learning using the same or different spatial information which we hypothesize will differ in content and underlying neurobiological mechanism. The miniscopes will be used to determine how the representations differ under changed conditions; 2) investigate effects of circadian rhythm perturbations on HPC function. The miniscope will be used to assess the impacts of various forms of circadian perturbations (i.e./ shift work) on HPC representations; and 3) memory function in the mammalian brain appears to be organized by neural networks mediating different memory functions and within this organization, normal thought and behaviour arise from cooperative and competitive interactions between these systems. We will explore the mechanisms of interactions amongst multiple memory networks using the miniscopes to image activity in subcortical and cortical regions under different experimental conditions. Adding in-vivo calcium imaging using UCLA-V4 will be highly beneficial to our trainees because: 1) optical imaging methods are becoming increasingly popular over traditional electrophysiological methods for various reasons including large amounts of data can be collected and yet less technical expertise is required to run and maintain the equipment; 2) making sense of the high volumes of data collected from the miniscopes requires computational skills which our trainees will receive; and 3) enhance the quality of publications generated. All of these aspects will be advantageous to our trainees as they pursue a competitive space in neuroscience. We will use this technology to advance our knowledge of how behaviour is shaped by exposure to specific events, the neural networks responsible, and the biochemical mechanisms supporting these functions. Our research will benefit all Canadians as we expand our understanding of normal and abnormal behaviour.